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Immunofluorescence Microscopy

Immunofluorescence Microscopy

A fluorescence microscope uses fluorescent chromophores called fluorochromes, which can absorb energy from a light source and then emit this energy as visible light. Fluorochromes include naturally fluorescent substances (such as chlorophylls) and fluorescent stains that are added to the specimen to create contrast. Dyes such as Texas red and FITC are examples of fluorochromes. Other examples include the nucleic acid dyes 4’,6’-diamidino-2-phenylindole (DAPI), and acridine orange.

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Related Experiment Video

Updated: Nov 9, 2025

Live Cell Imaging of Microtubule Cytoskeleton and Micromechanical Manipulation of the Arabidopsis Shoot Apical Meristem

Live Cell Imaging of Microtubule Cytoskeleton and Micromechanical Manipulation of the Arabidopsis Shoot Apical Meristem

Published on: May 23, 2020

An Optimized Whole-Mount Immunofluorescence Method for Shoot Apices.

Thu M Tran¹, Edgar Demesa-Arevalo¹, Munenori Kitagawa¹

¹Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, New York.

Current Protocols

|April 7, 2021

Summary

This summary is machine-generated.

This study presents a simple immunofluorescence protocol for visualizing protein localization in whole plant tissues like maize and Arabidopsis. The method aids in understanding protein functions and analyzing colocalization using ImageJ.

Keywords:

Arabidopsis immunofluorescence maize protein localization whole-mount

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Area of Science:

Plant Biology
Cell Biology
Molecular Biology

Background:

Protein localization is crucial for understanding biological functions.
Immunofluorescence is a key technique for protein visualization in cell biology.
A standardized protocol for whole-mount plant tissues is needed.

Purpose of the Study:

To describe an accessible immunofluorescence protocol for whole-mount protein localization in maize (Zea mays) and Arabidopsis.
To provide detailed steps and optimization tips for the protocol.
To offer a method for analyzing protein colocalization using ImageJ.

Main Methods:

Whole-mount immunofluorescence on maize ear primordia and Arabidopsis inflorescence apices.
Detailed protocol with step-by-step instructions and troubleshooting tips.
Application of the JACoP ImageJ plug-in for colocalization analysis.

Main Results:

An optimized protocol for whole-mount immunofluorescence in specific plant tissues (maize and Arabidopsis).
Demonstrated applicability for visualizing protein localization in 2-5 mm maize ear primordia and developing Arabidopsis inflorescence apices.
Successful colocalization analysis using the ImageJ plug-in.

Conclusions:

The developed protocol is effective for whole-mount protein localization in diverse plant tissues.
This method facilitates the study of protein function and interactions in plants.
The protocol serves as a valuable reference for plant biologists studying protein localization and colocalization.